CRYOPRESERVATION OF EJACULATED AND EPIDIDYMAL SPERM

目前在科学领域发生的遗传学革命正在为理解基因的生物学功能和阐明它们在人类疾病中的作用创造新的范例。这些类型的研究是有益的，因为它们可能产生新的诊断测试和治疗人类疾病的方法。突变小鼠的发生是建立基因功能及其在人类疾病中的作用策略的重要组成部分。因此，国际科学界产生的突变小鼠数量正以指数速度增长，而目前在储存库中储存这些小鼠的方法在成本和实用性方面并不理想。一个可能的解决方案是冷冻保存小鼠精子，但目前的方法是次优的和不一致的。本课题的目标是：(1)设计出最佳的小鼠精子射精方法，从这些研究者已经产生和研究的转基因、基因敲除和enu突变小鼠的野生型和特征良好的小鼠品系中射精；(2)对(1)所述小鼠品系的小鼠精子进行优化或开发新的冷冻保存方法；(3)将上述方法与宫内或输卵管内人工授精和/或体外受精方法和/或胞浆内精子注射相结合，以达到尽可能高的活产仔率；(4)用免疫表型法评价(3)活产小鼠的正常程度；组织病理学;行为基因表达（使用微阵列和/或SAGE分析）技术。

The genetics revolution currently occurring in the scientific arena is creating new paradigms in the understanding of the biological function of genes and elucidating their role in human disease. These types of studies are beneficial since they are likely to produce new diagnostic tests and treatments for human disease. The genesis of mutant mice is an important component of the strategy employed in establishing gene function and their role in human disease. Consequently, the number of mutant mice being generated by the international scientific community is increasing at an exponential rate and current methods of storing these mice in repositories are not ideal in terms of costs and practicality. One possible solution may be the cryopreservation of mouse sperm but current methodologies are sub-optimal and inconsistent. This proposal aims to: (1) devise optimal methods of ejaculating mouse sperm from wild-type and well-characterized strains of transgenic, gene knockout and ENU-mutagenized mice previously generated and studied by these investigators; (2) optimized/or develop new methods of cryopreservation of mouse sperm from strains of mice as mentioned in (1); (3) use the above methods in conjunction with intrauterine or oviductal insemination and/or in vitro fertilization methodologies and/or intracytoplasmic sperm injection so as to attain the highest possible rates of liveborn pups; and (4) evaluate the normality of liveborn mice from (3) using immunophenotyping; histopathology; behavioural gene expression (using microarray and/or SAGE analysis) techniques.